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Journal of Apiculture - Vol. 33 , No. 4

[ Original Article ]
Journal of Apiculture - Vol. 32, No. 4, pp.295-302
Abbreviation: J. Apic.
ISSN: 1225-0252 (Print)
Print publication date 30 Nov 2017
Received 27 Oct 2017 Revised 07 Nov 2017 Accepted 08 Nov 2017

Avoidance Behavior of Honey bee, Apis mellifera from Commonly used Fungicides, Acaricides and Insecticides in Apple Orchards
Moonsu Kang1 ; Chuleui Jung2, 3, *
1Korea Plants Environmental Research Station, Suwon, Republic of Korea
2Agriculture Science and Technology Research Institute, Andong National University, Republic of Korea
3Department of Plant Medicine, Andong National University, Republic of Korea

Correspondence to : * E-mail:

Funding Information ▼


Avoidance behavior is an important life history strategy to survive hazardous environment. The experiment was conducted to detect the avoidance tendency of the honeybee Apis mellifera against commonly used pesticides in apple production. Choice test given only 50% sucrose solution and pesticide-mixed sucrose solution as food estimated the avoidance in laboratory. Most of the acaricides and fungicides tested were shown avoided. Among insecticides, honeybee showed strong avoidance to cyhexatine, carbosulfan and fenpyroximate but low to diflubenzuron, tebufenpyrad, and acrinathrin. Avoidance behavior to neonicotinoid insecticides showed bifurcated; highly avoided from thiacloprid, acetamiprid while less avoided from imidacloprid, thiamethoxam and dinotefuran. From the field study, abamectin, fenthion, amitraz and acequinocyl showed highly avoided while fungicide of fenarimol, acaricides of acrinathrin and phosphamidon, IGR insecticide of diflubenzuron, neonicotinoid insecticide of imidacloprid, and carbamate insecticide of carbaryl showed less avoidance in the field. These results partly explained high bee poisoning from carbaryl in apple flowering period, and neonicotinoids during season.

Keywords: Apis mellifera L., Avoidance tendency, Pesticide, Selection index


Honey bees are vegetarians, usually consuming only nectar and pollen from plant blossoms or sweets like sugar syrup and honey dew (Atkin and Anderson. 1976 (1970)). These insects are highly social and a colony may contain as many as 10,000 to 100,000 individuals, depending on the time of year, prevailing weather conditions and availability of nectar and pollen sources (Robinson, 1979; Proctor et al., 1996). Honey bees provide not only the income sources for beekeepers but also pollination services for crop production (Degrandi-Hoffman, 1978; NRCS, 2005; Jung, 2008; Jung, 2014). Amongst beneficial insects, honey bees are considered as the most efficient and reliable pollinators of various agricultural crops (McGregor, 1976). Recently increased concerns on honey bee population decline have wide spread not only within Korea but over the world (Jung, 2014). There are several factors contributing to the decline of honey bees as well as the pollination services (Goulson et al., 2015), but considered as the combined stresses from parasites, pesticides and lack of foraging sources. Habitat destruction, including nesting and mating sites, and alternative forage, is the main issue in the decline of pollinators (Batra, 1995). Bee poisonings from diverse chemicals especially of insecticides are another important source of honeybee population decline together with parasitic pests and diseases (Crane and Walker, 1983; Choi and Lee, 1986). Pesticides constitute an important component of modern agriculture without which the increase in agricultural production could never be achieved (Atkins et al., 1970). However, the large scale and indiscr-iminate use of pesticides has resulted in great conflict of interests because of simultaneously occurrences of harmful effects to biotic and abiotic agricultural environments (Kevan and Plowright, 1995). Many agrochemicals sprayed in the fields for crop protection against insect and disease pests are injurious to honeybees (Johansen, 1977). Impacts of these chemicals can occur directly as the mortality on contact or ingestion or indirectly elicit behavioral disorder or abnormal development of immatures. In many cases of acute toxicity, poisoned bees often become irritable (likely to sting), paralyzed or stupefied, appear to be ‘chilled’ or exhibit other abnormal behavior (Dharmawardena, 1994).

Avoidance behavior is one of the behavioral survival mechanisms of organism together repair or tolerance of molecular or physiological damage from toxic chemicals (Bowler et al., 1992; Li-Byarlay et al., 2016), Avoidance behavior includes irritancy which is occurring after physical contact and repellency occurring without physical contact but often with olfactory means (Chareonviriyaphap et al., 1997). Here we assessed the avoidance tendencies of relatively low toxic pesticides such as acaricides and fungicides commonly used in apple production in Korea. Choice test given only 50% sucrose solution and pesticidemixed sucrose solution as food estimated the avoidance in laboratory and field assessments were measured after spraying each selected chemical.

Pesticide avoidance test in the laboratory

Pesticides: Sixty three pesticides (Table 1, 2, 3; 16 fungicides, 12 acaricides and 35 insecticides) were tested for the study. From the registry of pesticides on apple, selection was based on the chemicals that could be used in blooming season as the first priority and then mostly common use as the second priority.

Honeybee: Honey bees used in the experiments were from the experimental apiary of Andong National University. About 30 colonies of Apis mellifera, hybrids of A. m. ligustica were maintained with conventional methods but limited pesticide uses. For the avoidance assay, mixed ages of worker bees were collected from randomly chosen hives.

Cylindrical cages made of stainless wire ( 13.5 x H10.5cm) were used for this test. Five honey bees were kept in one cage considered as single replication and total 5 replications per pesticide were conducted. No food or water was provided for 2 hours before the experiments. Then, 50% sucrose solution and pesticide-mixed 50% sucrose solutions retaining recommended dose of pesticides were provided into two separate pots maintaining 9cm distance for detecting pesticide avoidance under 25±3°C, 50~80% RH and dark condition. Choice of each food source was checked for 5 minutes at the intervals of 1, 2 and 3 hours. When bee is on the food item and showed feeding behavior of touching food and retracting proboscis, it was considered as choice. The avoidance index was calculated by the formula:

Table 1. 
List of fungicides with chemical class, common name, amount of active ingredient (AI, %), formulation type and recommended concentration (g or ml of formulated product/20L) used in the laboratory test and in the field (**)
Class Common name AI% Type R.C.
Surfactant Lime sulfur 22 Ls 200.0
Guanidine Iminocatadine triacetate 3 Ec 13
Dicarboxmide Iprodione 50 Wp 17
Benzimidazole Benomyl 50 Wp 13
Anilinopyrimidine Pyrimethanil 30 Wp 20
Dithiocarbamate Mancozeb 75 Wp 40
Dithiocarbamate Propineb 70 Wp 40
Carbamate Carbendazim 60 Wp 20
Triazole Diniconazole 5 Wp 10
Triazole Difenoconazole 10 Wp 6.7
Triazole Myclobutanil 6 Wp 13
Triazole Bitertanol 25 Wp 20
Triazole Imibenconazole 15 Wp 10
Triazole Flusilazole 2.5 Wp 2.5
Pyrimidine Fenarimol ** 12 Wp 6.7
Antibiotic Polyoxin b 10 Wp 20

Table 2. 
List of acaricides with chemical class, common name, amount of active ingredient (AI, %), formulation type and recommended concentration (g or ml of formulated product/20L) used in the laboratory test and in the field (**)
Class Common name AI% Type R.C.
Naphthoquinone Acequinocyl ** 15 Sc 20
Amidine Amitraz ** 20 Ec 20
Triflurea Flufenoxuron 5 Sc 20
Sulfite ester Propargite 30 Wp 27
Organochlorine Tetradifon 8 Ec 25
Organophosphate Fenthion ** 50 Ec 20
Organophosphate Phosphamidon ** 50 Sl 20
Organotin Azocyclotin 25 Wp 13
Organotin Fenbutatin oxide 50 Wp 20
Carbazate Bifenazate 23.5 Sc 10
Tetronic acid Spirodiclofen 36 Wp 10
Machine oil Machine oil 98 Ec 100

Table 3. 
List of insecticides with chemical class, common name, amount of active ingredient (AI, %), formulation type and recommended concentration (g or ml of formulated product/20L) used in the laboratory test and in the field (**)
Class Common name AI% Type R.C.
Benzoylurea Diflubenzuron ** 25 Wp 8
Diacylhydrazine Tebufenozide 8 Wp 20
Organophosphate Trichlorfon 50 Sl 25
Organophosphate Fenitrothion 50 Ec 25
Organophosphate Monocrotophos 24 Sl 20
Organophosphate Acephate ** 50 Wp 20
Organophosphate Azinphos methyl 25 Wp 40
Organophosphate Chlorpyrifos 25 Wp 20
Organophosphate Parathion 17 Ec 20
Organophosphate Dichlorvos ** 50 Ec 20
Organophosphate Epn 45 Ec 20
Organotin Cyhexatin 25 Wp 13
Carbamate Carbaryl ** 50 Wp 25
Carbamate Methomyl ** 45 Wp 13
Carbamate Carbosulfan 20 Wp 20
Neonicotinoid Acetamiprid 8 Wp 10
Neonicotinoid Imidacloprid ** 10 Wp 10
Neonicotinoid Thiacloprid 10 Sc 10
Neonicotinoid Thiamethoxam 10 Wp 10
Neonicotinoid Clothianidin 8 Gr 10
Neonicotinoid Dinotefuran ** 10 Wp 20
Phenoxypyrazole Fenpyroximate 5 Sc 10
Pyrazole Tebufenpyrad 10 Ec 10
Pyridazinone Pyridaben 20 Wc 20
Pyrethroid Alpha cypermethrin 2 Ec 20
Pyrethroid Deltamethrin 1 Ec 20
Pyrethroid Bifenthrin 2 Wp 20
Pyrethroid Acrinathrin ** 3 Wp 6.7
Pyrethroid Esfenvalerate ** 1.5 Ec 20
Pyrethroid Etofenprox 10 Wp 20
Pyrethroid Fenpropathrin 5 Ec 20
Pyrethroid Cypermethrin ** 5 Ec 20
Pyrethroid Lambda cyhalothrin 1 Wp 20
Antibiotic Milbemectin 1 Ec 20
Antibiotic Abamectin ** 1.8 Ec 6.7

where AI = avoidance index, Np = number of bees chosen pesticide-mixed sucrose solution, and Nc = number of bees chosen sucrose solution. The lower the avoidance index (AI) value, the higher the avoidance tendency. If the value is significantly lower than 0.5, it was considered as avoiding.

Pesticide avoidance test in the field

Field trial was conducted at the apple orchard in Geumgok, Giran-myeon, Andong, Korea. Main cultivar is Fuji with conventional M-26 rootstock. 2-3m high apple trees were lined with 3m apart each other. The bee species A. mellifera cultured in Andong National University was used in this test. One fungicide (fenarimole), four acaricides (acequinocyl, amitraz, fenthion, phosphamidon), and 11 insecticides (diflubenzuron, acephate, dichlorvos, carbaryl, methomyl, imidacloprid, dinotefuran, acrinathrin, esfenvalerate, alpha cypermethrin, abamectin) were applied for this study (Table 1,2,3 with ** mark). Selection was made based on the toxicity data in the laboratory as well as the prevalent use in apple orchards (Kang, 2009).

Each pesticide was applied to branches of the tree. Pesticide was sprayed using the knapsack sprayer two times, 7 and 1 day prior to full blooming using the recommended dose with application rate of 400L per hectare. Then, three trees on which branches all 16 pesticides were sprayed were caged by using the nylon net (10x4x5m), after last spray. The one cage was considered as replicate. Then, one honeybee colony with queen right containing ap. 1500 workers was introduced into each cage in 1 day after final spray. Net was used to protect the honey bees from escaping. Numbers of honey bees visiting the flower blossoms were counted for 10 minutes for 2,3,4 DAT (day after treatment) and 8,9,10 DAT, respectively around noon.

Data analysis

Avoidance data from the laboratory study were analyzed by one tailed t-test with mean equal 0.5 using SAS program (SAS Institute, 2004). Avoidance index data from the field were compared by ANOVA, Turkey test. Honeybee visiting data to flower in the field between two time laps were compared with t-test.

Pesticide avoidance test in the laboratory

In this study, honeybee showed significant avoidance to all tested fungicides (Fig. 1, P<0.05) in the laboratory test. This means that honeybee discriminate fungicide-mixed sucrose solution. Only to iminocatadine triacetate, the AI values was marginally significant (P=0.07). This chemical is used for apple bitter rot or valsa canker management. AI index of most fungicides used in apple orchards showed ranges from 0.12 to 0.37 with carbendazim lowest. Carbendazim is relatively non-toxic to honeybee in contact and feeding test (Kang, 2009).

Fig. 1. 
Avoidance indices (Mean±SE) of honeybees to fungicides. Value significantly lower than 0.5 means high avoidance.

Most of acaricides tested in the experiment showed lower than 0.2 AI values indicating significant avoidance tendency (Fig. 2). AI value of phosphamidon was not significant which means honeybee does not discriminate the chemical (P=0.24). Phosphamidon is an organophosphate insecticide developed in 1960s. It acts as a choline esterase inhibitor, and showed high toxicity to honeybee in the laboratory experiments (Kang, 2009), while the lowest avoidance index was observed from acequinocyl (0.07), which showed very low toxicity to honeybee in the contact and feeding test (Kang, 2009).

Fig. 2. 
Avoidance indices (Mean±SE) of honeybees to acaricides. Value significantly lower than 0.5 means high avoidance.

Honeybees showed significant avoidance to most of the insecticide tested (Fig. 3, P<0.05), except chlorpyrifos (P=0.06), tebufenpyrad (P=0.07) and diflubenzuron (P=0.17). The avoidance index was found lowest (0.09) in cyhexatin and highest in diflubenzuron (0.37).

Fig. 3. 
Avoidance indices (Mean±SE) of honeybees to insecticides. Neonicotinoid insecticides were marked with *. Value significantly lower than 0.5 means high avoidance.

Interestingly, avoidance behavior to neonicotinoid insecticides showed bifurcated patterns; highly avoided (AI<1.2) to thiacloprid, acetamiprid while less avoided (AI>2.5) to thiacloprid, imidacloprid, thiamethoxam and dinotefuran. Imidacloprid and thiaclopriod are much more toxic to honeybee than acetamiprid or thiacloprid in the laboratory contact or feeding trials or field, leading EU ban of those chemicals (Cressey, 2013; Lee et al., 2016). Less discrimination of these chemicals by honeybee could result in synergized toxicity in the field to honeybee (Woodcock et al., 2016; Alburaki et al., 2017) or bumblebee (Arce et al., 2017). If bees are exposed to guttation of the plants where neonicotinoids were applied into the soil such as seed coating, the negative impact could exacerbated by the increased concentration of the systemic pesticide (Girolami et al., 2009).

Pesticides avoidance test in the field

From the field trials evaluating avoidance tendency of honeybee from pesticide-treated flower blossoms, flower visiting rates were variable on treatments. Fenarimol, diflubenzuron and carbary showed no difference on the flower visiting rates of honey bees measured 2-4 DAT and 8-10 DAT (Fig. 4). This implies that those chemicals would not elucidate the behavioral changes of honeybee foraging possibly via contact or olfactory cures. Flower visiting rates were relatively lower in acequinocyl, amitraz, fenthion, acephate, diclorovos and abamectin. All treatments showed avoidance. AI values were ranged from 0.23 to 0.42 (Fig. 4). The responses were categorized into three groups; highly, moderately and marginally avoided. Abamectin, fenthion, amitraz and acequinocyl showed highly avoided which showed high avoidance in the laboratory test, too. While fungicide of fenarimol, acaricides of acrinathrin and phosphamidon, IGR insecticide of diflubenzuron and neonicotinoid insecticide, imidacloprid showed less avoidance in the field test. Interestingly, fewer numbers of honeybees were foraging abamectin and fenthion sprayed flowers, but high numbers of honeybees were found forging on flowers where difulbenzuron and carbaryl were sprayed (Fig. 4). Recently more apple growers spay carbaryl for flower and fruit thining, higher toxic effects on foraging honeybee as well as broods are expected. This result could partly explain the massive bee poisoning during apple flowering season. Kim et al. (2014) reported that carbaryl is the most responsible chemical imposing greater risk to honeybee during apple flowering season.

Fig. 4. 
The comparison of visiting rate of honeybees to the apple flowers at 2 to 4 and 8 to 10 days after spray. Sixteen pesticides were sprayed and flower visiting rates (No. bees/blossoms/10 min) were estimated. On the same chemical, different letters on the bar indicate significant difference of visiting rates in two-time frames.

Fig. 5. 
Avoidance tendency of honeybees from the flower blossoms where 16 pesticides were sprayed in apple orchards.

Avoidance is an adaptive behavior through which an organism could escape the hazard for better survival. Through the series of laboratory and field experiments, we found that honeybee showed significant avoidance tendencies from the toxic pesticides. Interestingly, avoidance tendencies were much strong to fungicides and acaricides than to insecticides. Lower physiological susceptibility to fungicides or acaricides together with strong avoidance would benefit to survival of honeybees. But weaker avoidance to some insecticides including neonicotinoids of imidacloprid or thiamethoxam could lead to danger of bee poisoning.


This work was partly supported from RDA agenda project (PJ01303701) on evaluation of honey pollination services for crop production.

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